Large numbers of amplicons or polymerase chain reaction products (PCR) products are necessary to amplify every exon in a genome. For example, the amplification all of the exons in the human genome would require approximately 250,000 amplicons or PCR products. For each amplicon, a pair of primers needs to be synthesized and an individual PCR reaction needs to be carried out, which is a costly and unwieldy task for the total number of 250,000 amplicons required to sequence all of the exons in the human genome.
Microarray technologies are available that can synthesize very small quantities of up to about 400,000 oligonucleotides on a single solid support. These 400,000 oligonucleotides could potentially comprise 200,000 primer pairs for an amplification reaction, such as PCR. All 400,000 oligonucleotides can be liberated from the microarray into a single (multiplex) PCR reaction using known methods. However, the resulting multiplex PCR using these 200,000 primer pairs in one reaction is likely to be uninformative due to the complexity of the reaction and due to numerous artifacts, such as primer-dimer formation. There is currently no known method to segregate individual primers pairs from a microarray into individual PCR reactions.
Thus, a need exists to segregate primer pairs from a microarray, such that individual primer pairs can be used, for example, in an amplification reaction.